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Determinants of RNA Integrity and Purity


RNA quality is a critical determinant for the success of many downstream applications, including microarray analysis. For this reason, if you are planning microarray analysis experiments, it is important to fully consider the factors that affect the quality of your RNA.


Tissue-Specific Responses to Injury Affect RNA Integrity
Traditionally, efforts to preserve RNA quality have focused on methods of tissue storage and disruption, with the goal of minimizing RNase activity. However, a more critical determinant is actually the RNA quality within tissues before the RNA expression pattern is 'frozen' by preservation. RNA integrity within the cell is dependent on a complex series of responses that are set in motion in response to insult, as well as the interval between the time of injury and tissue preservation. The mechanisms of RNA degradation can be both complex and varied, as is the resulting impact on the mRNA population. Therefore, no matter how carefully you prepare your RNA, the integrity is often determined before the tissue sample reaches your hands.


Residual Contaminants -- The Hidden RNA Quality Factor
Even the most intact RNA will not perform well if the sample contains trace contaminants. The most detrimental contaminants are residual organics, metals, and proteins such as nucleases. RNA that contains these impurities will perform poorly in most enzymatic applications.
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